Hard capsules

ABSTRACT

The present disclosure concerns embodiments of new hard pullulan capsule shells and capsules comprising (I) moisture, (II) a mono-, di-, and oligosaccharides free pullulan and (III) a setting system. Also provided are an aqueous composition and a dip-molding manufacturing method for the manufacture of such shells and capsules.

This application is national phase application based on InternationalApplication No. PCT/IB2012/000176, filed Jan. 10, 2012 which claims thebenefit of U.S. Provisional Application No. 61/431,496, filed Jan. 11,2011, all of which are incorporated herein by reference on theirentirety.

FIELD OF THE INVENTION

The present invention relates to new hard capsule shells and capsulescomprising pullulan, a dip-molding process for the manufacture thereof,aqueous dispersions for use in the dip-molding process and the use of aspecific pullulan grade for the manufacture of hard capsule shells andcapsules.

BACKGROUND OF THE INVENTION

Hard and soft capsules comprising pullulan are known.

WO2005105051 discloses a hard capsule comprising a conventional pullulangrade in an amount of 85% to 90% by weight, potassium chloride in anamount of 1.0% to 1.5% by weight, carrageenan in an amount of 0.1% to0.4% by weight, one or more surfactants in an amount of 0.1% to 0.2% byweight and water in an amount of 10% to 15% by weight over the weight ofthe capsule. Thanks to the addition of a particularly selected settingsystem, WO2005105051 capsules are inter alia endowed with an improvedchemical stability (e.g. no polymer cross-linking as with conventionalgelatin). WO2005105051 does not disclose or suggest the use of anyspecific pullulan grade or the advantages following therefrom.

EP1072633 discloses compositions based on pullulan and a setting systemfor the use e.g. in manufacturing soft or hard capsules. EP1072633stresses the importance of the addition of an appropriate setting systemwhereas no indication is given to select a specific pullulan grade orthe advantages following therefrom.

EP1157691 discloses pullulan compositions for use e.g. in manufacturingsoft or hard capsules with improved surface properties. The advantage isobtained by using a surfactant of pharmaceutical or food quality.EP1157691 does not disclose or suggest the use of any specific pullulangrade or the advantages following therefrom.

EP1398346 discloses the manufacturing of a high pullulan content shapedproducts (such as hard capsules) comprising α,α-threalose and a pullulanhaving an average molecular weight between 20,000 and 4,000,000 Dalton,preferably between 50,000 and 2,000,000 Dalton, in an amount of 30% orhigher on a dry solid basis. The presence of α,α-threalose is praised asthe key factor to impart the final products with properties such asstability to the change of humidity, transparency, gloss, and solubilityin water. Although the use of PI-20, PF-10 and PF-20 pullulan grades isdisclosed, no indication is given to select other specific and moreadvantageous pullulan grades.

JP5-65222-A describes a soft capsule, capable of stabilizing a readilyoxidizable substance enclosed therein, exhibiting easy solubility, andbeing able to withstand a punching production method. The soft capsuleis obtained by blending a capsule film substrate such as gelatin, agar,or carrageenan with pullulan. No indication is given to select aspecific pullulan grade or the advantages following therefrom.

The hard capsule shells disclosed in the prior art present certaindrawbacks, notably a non fully satisfactory shell mechanical strength(i.e. shell brittleness) at shell low LOD. Improving this property is aparticularly desirable goal for pullulan hard capsule shells. Pullulanis in fact more sensitive than other materials (e.g. gelatin or HPMC) tolow moisture conditions. This sensitivity impinges upon increased shellbrittleness at low water content. Unsatisfactory brittleness meanshigher manufacturing losses, a poorer quality and global higher costs.On the other hand, reducing LOD of the shell might be desirable toencapsulate those active principles and natural or synthetic substancesthat are sensitive to water. A shell low LOD is also obtained when hardshells are filled with hygroscopic substances that make the shellmoisture to migrate from the shell to the filling so as to simulate afurther drying of the shell.

Accordingly, one object of the present invention is therefore theprovision of pullulan hard capsule shells and capsules which overcomethe drawbacks of the prior art. In particular, one object of the presentinvention is the provision of new pullulan hard shells and capsules thatwith respect to prior art pullulan hard capsule shells present improvedcharacteristics in terms of shell mechanical properties especially atlow shell LOD levels, and an equally good or improved shell dissolutionprofile, shell transparency, capsule surface static and glidingproperties, absence of unpleasant smell, capsule compatibility withautomatic high speed capsule filling equipment.

Another object of the invention is to provide a dip-moldingmanufacturing process that allows for the manufacture of the improvedhard capsule shells as above, in an easy and cost-effective manner andthat allows maintaining capsule dimension specificities duringproduction.

SUMMARY OF THE INVENTION

The above and other objects are achieved by an aqueous composition forthe manufacture of hard capsule shells characterized in that itcomprises (i) an aqueous solvent, (ii) a mono-, di-, andoligosaccharides free pullulan and (iii) a setting system.

The above and other objects are also achieved by a hard capsule shellcomprising (I) moisture, (II) a mono-, di-, and oligosaccharides freepullulan and (III) a setting system.

The above and other objects are also achieved by a hard capsulecomprising a shell as defined above filled with one or more substancesin solid, semi-solid and/or liquid form.

The above and other objects are also achieved by a dip-molding methodfor the manufacture of hard capsule shells as defined above, saidprocess comprising the steps of:

(a) dipping pins into an aqueous composition as defined above

(b) withdrawing the dipping pins from the aqueous composition and

(c) drying the composition adhered on the dipping pins so as to obtain ashell;

wherein the steps (a) to (c) are performed in the order in which theyare presented.

The above and other objects are also achieved by the use of a mono-,di-, and oligosaccharides free pullulan for the manufacture of hardcapsule shells.

FIGURES

FIG. 1 is a graph representing shell mechanical properties for shells ofthe present disclosure (“Sample #7” of Example 4) and comparativeconventional pullulan shells (“Sample #8” of Example 4) by means of thetube test after storage at different RH conditions, illustrating thatthe comparative conventional pullulan shells exhibit higher brittlenessas compared to the shells of the present disclosure, wherein “Broken %”refers to the percentage of broken capsules and “LOD %” refers toloss-on-drying;

FIG. 2 is a graph representing shell transmittance measurements forshells of the inventions (Sample#7—see sample details in example 4) and(comparative) conventional pullulan shells (Sample#8—see sample detailsin example 4). Transmittance measured with spectrophotometer over thewavelength from 400 to 700 nm;

FIG. 3 is a graph representing shell dissolution properties for shellsof the inventions (Sample#7—see sample details in example 4—identifiedby Lot#166181) and (comparative) conventional pullulan shells(Sample#8—see sample details in example 4—identified by Lot#165651).Shells were filled with APAP. Test conditions are disclosed below.Results in the graph are the average of the values obtained for 6capsules.

DETAILED DESCRIPTION OF THE INVENTION

Below, any consideration and embodiment disclosed in connection with oneaspect of the invention (e.g. the aqueous composition) must beunderstood to equally apply to the other aspects of the invention (e.g.hard capsule shells, hard capsules, the dip-molding process and uses) tothe extent that it is technically possible.

In a first aspect, the present invention relates to an aqueouscomposition for the manufacture of hard capsule shells characterized inthat it comprises (i) an aqueous solvent, (ii) a mono-, di-, andoligosaccharides free pullulan and (iii) a setting system.

In one embodiment, the aqueous composition of the invention has a totalamount of solids typically comprised between about 10% and 55% byweight, preferably between about 15% and 40%, more preferably between20% and 30% by weight over the total weight of the composition.

In one embodiment, the aqueous composition of the invention has a totalamount of solids so that the viscosity of the aqueous composition at atemperature comprised between about 50° C. and 60° C. is comprisedbetween about 500 cPs and 1500 cPs, preferably between about 800 cPs and1000 cPs.

Unless otherwise indicated, the term “solids” includes at least all nonaqueous ingredients added into the aqueous composition of the invention.Solids in particular comprise a mono- di, and oligosaccharide freepullulan and a setting system. Solids also comprise other optionalingredients typically used in hard capsule manufacturing such asplasticizers, surfactants, sequestring agents, pharmaceutically or foodacceptable flavouring agents, pharmaceutically or food acceptable dyesand pharmaceutically or food acceptable colouring agents, if present.

Unless otherwise indicated, “aqueous solvent” preferably means water,more preferably deionised water. More preferably “aqueous solvent”consists of deionised water. In one embodiment, demineralised water ispreferably demineralised water for pharmaceutical uses as defined underUSP purified water in the USP-32.

Pullulan (CAS number 9057-02-7; chemical synonyms 1,4-1,6-α-D-Glucan,1,6-α-linked maltotriose) is a polysaccharide polymer consisting ofmaltotriose units. Three glucose units in maltotriose are connected byan α-1,4 glycosidic bond, whereas consecutive maltotriose units areconnected to each other by an α-1,6 glycosidic bond. Maltotetraose unitsconsisting of four 1,4-linked glucose molecules also occur, probablyrandomly, but are rare (about 6%). There is also evidence for a rareoccurrence of branching points where poly-maltotriosyl side-chains areattached to the main chain by a 1,3-glycosidic bond.

Pullulan is generally produced from starch by the fungus Aureobasidiumpullulans. Pullulan has good film forming properties and a particularlylow oxygen permeability. Its existence was reported for the first timein 1938. Hayashibara Company started the commercial production in 1976.

Unless otherwise indicated, pullulan of the present invention is apullulan as defined above which is further characterized by being freeof mono-, di-, and oligosaccharides. Pullulan can be made free of mono-,di-, and oligosaccharides either throughout a post-manufacturing processstep (e.g. precipitation of the mono-, di-, and oligosaccharides from asample of a commercial pullulan grade in ethanol) or from its veryproduction, by choosing appropriate starch fermentation conditions.Mono-, di-, and oligosaccharides free pullulan for use in this inventionhas been purchased from Hayashibara Company, Okayama Japan.

In one embodiment, the pullulan of the present invention has preferablyan average molecular weight comprised between about 100 KDa and 400 KDa,preferably between 150 KDa and 350 KDa, more preferably between 200 KDaand 330 KDa.

In one embodiment, the pullulan of the present invention has preferablya melt viscosity at about 60° C. between about 500 cPs and 1500 cPs,preferably between about 800 cPs and 1000 cPs.

In one embodiment, “mono-, di-, and oligosaccharide free pullulan”preferably means that the amount of mono-, di- and oligosaccharides in apullulan sample is preferably lower than 2.5%, preferably lower or equalto 1%, preferably lower than or equal to 0.9%, more preferably lowerthan or equal to 0.7%, even more preferably lower than or equal to 0.6%,by weight over the total weight of the pullulan sample, on a dry solidbasis. In one embodiment, “dry solid basis” preferably means that thecontent of mono-, di-, and oligosaccharides is measured in a sample ofpullulan containing water in an amount comprised between about 0.5% and2% by weight over the total weight of the sample.

Unless otherwise indicated, mono-, di-, and oligosaccharides of pullulanare measured as glucose units. Unless otherwise indicated, a pullulanmonosaccharide is preferably a glucose monomer; a pullulan disaccharideis preferably a glucose dimer; a pullulan oligosaccharide preferablycomprises between about 3 and 10 glucose monomers.

Mono-, di-, and oligosaccharides are the main pullulan impurities.Methods to calculate the % by weight of mono-, di-, and oligosaccharides(as glucose units) over the total weight of a pullulan sample are knownto a skilled man. For example, reference can be made to theanthrone-sulfuric acid method disclosed in Morris D. L. (1948)“Quantitative determination of carbohydrates with Dreywood's anthronereagent”, Science 107: 254-255:

Equipment

Spectrophotometer capable of measuring absorbance at 620 nm

Procedure

Preparation of standard: Weigh accurately 0.2 g glucose, dissolve inwater and make up to 1 l.

Measurement of mono-, di- and oligosaccharides

Weigh accurately 0.8 g sample and dissolve in water to make 100 ml(stock solution). Place 1 ml of the stock solution in a centrifuge tube.Add 0.1 ml saturated potassium chloride solution. Add 3 ml methanol andmix vigorously for 20 sec. Centrifuge at 11000 rpm for 10 minutes. Add0.2 ml of the supernatant to 5 ml modified anthrone solution (0.2 ganthrone in 100 g 75% (v/v) sulfuric acid, freshly prepared). Add 0.2 mlof glucose standard solution and 0.2 ml water (blank control) toseparate 5 ml portions of modified anthrone solution. Mix rapidly. Placesamples in a 90° water bath and incubate for 15 min. Measure absorbanceof the test solution at 620 nm. Calculate the percent of mono-, di- andoligosaccharides expressed as glucose (C) in the sample:C(%)=[(At−Ab)×0.41×G×100]/(As−Ab)×W

where

At is absorbance of the test solution

Ab is absorbance of the water blank

As is absorbance of the standard solution

G is weight of the glucose (g)

W is weight of the sample (g)

Without wanting to be bound by any theory, it is believed that the useof a mono-, di-, and oligosaccharides free pullulan plays an importantrole in achieving the improved properties of the shells and capsules ofthe invention, notably in terms of improved shell mechanical propertiesespecially at low shell LOD levels. A commercially available pullulangrade is for example Pullulan PI-20 (tradename from Hayashibara) where“P” stands for “pullulan”, “I” for “de ionized” and the FIG. 20designates the average molecular weight of about 200 kDa. Anothercommercially available de ionized pullulan grade is PI-10 (tradenamefrom Hayashibara) having average molecular weight of about 100 kDa. Foodgrade pullulans are also commercially available. For example,Hayashibara commercializes PF-10 and PF-20 grades (both tradename)having average molecular weights of about 100 kDa and 200 KDa,respectively. Typically, a sample of a commercially available pullulangrade (such as grade PI-20) contains between 7% and 10% by weight ofmono-, di- and oligosaccharides over the total weight of pullulansample, on a dry basis. As shown in the examples of the presentapplication, these levels of mono-, di- and oligosaccharides do notallow obtaining the advantageous properties shown by the hard capsuleshells and capsules of the invention.

In one embodiment, the aqueous composition of the invention contains amono-, di-, and oligosaccharides free pullulan as defined above in anamount suitable to obtain a hard capsule shell as defined belowcontaining between about 65% and about 99% by weight, preferably betweenabout 65% and about 98% by weight, more preferably between about 70% andabout 97% by weight, more preferably between about 85% and about 96% byweight of such pullulan over the weight of the shell. Hard capsuleshells meeting the above weight % can typically be manufactured viadip-molding by using aqueous compositions comprising between about 15%and 40%, preferably between about 15% and 30%, even more preferablybetween about 18% and 25% by weight of such pullulan over the totalweight of the aqueous composition.

In one embodiment, the mono-, di-, and oligosaccharides free pullulan asdefined above represents more than 50%, preferably more than 75%, evenmore preferably 100% by weight over the weight of the total pullulanpresent in the aqueous composition, the shells and capsules of thepresent invention.

In one embodiment, the mono-, di-, and oligosaccharides free pullulan isproduced from a generic source of starch, preferably the starch is cornor tapioca starch. In one embodiment, the mono-, di-, andoligosaccharides free pullulan is produced from corn starch. In oneembodiment, the mono-, di-, and oligosaccharides free pullulan isproduced from tapioca starch.

In one embodiment, the aqueous composition of the invention optionallycomprises additional film-forming polymers typically used in themanufacture of hard capsule shells. Typically, such additionalfilm-forming polymers are selected from the group consisting of:gelatin, polyvinyl alcohol, starch, starch derivatives (for examplehydroxyethylated and hydroxypropylated starches), cellulose, cellulosesderivatives (for example HEC, HMC or HPMC such as HPMC USP30-NF25 grade2208, 2906 or, preferably, 2910) and mixtures thereof. Preferably,additional film-forming polymers are selected from the group consistingof: gelatin, HPMC, starch derivatives and mixture thereof. In oneembodiment, the aqueous composition of the invention optionallycomprises gelatin. In one embodiment, the aqueous composition of theinvention optionally comprises HPMC. In one embodiment, the aqueouscomposition of the invention optionally comprises starch derivatives.

In one embodiment, the mono-, di-, and oligosaccharides free pullulan asdefined above represents more than 50%, preferably more than 75%, evenmore preferably 100% by weight over the weight of all the film-formingpolymers typically used in the manufacture of hard capsule shells andoptionally present in the aqueous composition, the shells and capsulesof the present invention. Preferably, such film-forming polymers are asdefined above. The embodiment wherein the pullulan of the inventionrepresents 100% by weight of all the film-forming polymers present inthe aqueous composition preferably means that the aqueous composition ofthe invention contains traces of, or more preferably does not containany one of the following polymers: gelatin, polyvinyl alcohol, starch,starch derivatives (for example hydroxyethylated and hydroxypropylatedstarches), cellulose, celluloses derivatives (for example HEC, HMC orHPMC such as HPMC USP30-NF25 grade 2208, 2906 or preferably 2910) andmixtures thereof.

Setting systems are conventionally relied upon in the manufacture ofhard capsule shells by non-thermogelling dip-molding processes to conferan appropriate setting ability with cooling to film-forming polymers(like pullulan, HPMC or starch derivatives) that in these conditionshave per se poor gelling properties. The setting system makes theaqueous composition to set on the dipped pins, thus assuring a uniformcapsule shell thickness.

A huge patent literature provides guidance as to select the mosteffective setting system(s) depending on the film-forming polymers to beused in the capsule shell manufacturing. Reference can be made forexample to U.S. Pat. No. 5,264,223 and EP714656 (discussing HPMCcapsules), EP1117736 (discussing starch derivatives capsules);WO2005105051 and EP1072633 (discussing pullulan capsules).

In one embodiment, the setting system of the invention comprises one ormore gelling agents. In one embodiment, the setting system of theinvention comprises one or more gelling agents and one or more gellingaids (also known as co-gelling agents).

In one embodiment, the one or more gelling agents are selected from thegroup consisting of alginates, agar gum, guar gum, locust bean gum(carob), carrageenan (preferably kappa and/or iota), tara gum, arabicgum, ghatti gum, khaya grandifolia gum, tragacanth gum, karaya gum,pectin, arabian (araban), xanthan, gellan gum, starch, konjac mannan,galactomannan, funoran, acetan, welan, rhamsan, furcelleran,succinoglycan, scleroglycan, schizophyllan, tamarind gum, curdlan,dextran and mixtures thereof. Preferably, the one or more gelling agentsare selected from the group consisting of carrageenans (preferably kappaand/or iota, more preferably kappa-carrageenans), gellan gum andmixtures thereof. In one embodiment, the one or more gelling agentscomprise, preferably consist of carrageenans (preferably kappa and/oriota, more preferably kappa-carrageenans). In one embodiment, the one ormore gelling agents comprise, preferably consist of gellan gum. In oneembodiment, the one or more gelling agents comprise a combination of twoor more of the agents listed above. In one embodiment, the one or moregelling agents comprise, preferably consist of a combination of xanthanand locust bean gum. In one embodiment, the one or more gelling agentscomprise, preferably consist of a combination of xanthan with konjacmannan.

In one embodiment, the one or more gelling aids (also known asco-gelling agents) are cations. In one embodiment, the one or moregelling aids are selected from the group consisting of: K⁺, Li⁺, Na⁺,NH₄ ⁺, Ca²⁺, Mg²⁺ and mixtures thereof. Preferably, the one or moregelling aids are selected from the group consisting of: K⁺, NH₄ ⁺, Ca²⁺and mixtures thereof. The cations can be added to the setting system inthe form of a pharmaceutically or food acceptable water soluble salt(e.g. chloride, citrate or phosphate).

In one embodiment, the setting system of the invention comprises,preferably consists of:

-   -   one or more gelling agents selected from the group consisting of        carrageenans (preferably kappa and/or iota, more preferably at        least kappa-carrageenans), gellan and mixtures thereof; and    -   one or more pharmaceutically or food acceptable water soluble        salts of K⁺, NH₄ ⁺, Ca²⁺ and mixtures thereof.

In one embodiment, the aqueous composition of the invention contains oneor more gelling agents as defined above in an amount suitable to obtaina hard capsule shell as defined below containing between about 0.01 and3.0%, by weight, preferably between about 0.03 and 1.0%, by weight,preferably between about 0.1% and 0.5% by weight of such gellingagent(s) over the weight of the shell. Exemplary suitable gellingagents' amounts are readily available to a skilled person in the fieldof hard capsules manufacturing. For example it is commonly accepted thathard capsule shells containing a “target” amount of gelling agentsfalling within the ranges identified above can be obtained via adip-molding process by using aqueous compositions containing about ¼(i.e. 25%) of that target amount (expressed as % by weight over theweight of the composition).

In one embodiment, the aqueous composition of the invention contains oneor more gelling aids as defined above in an amount suitable to obtain ahard capsule shell as defined below containing about less than 3%,preferably about less than 2.0%, more preferably between about 0.5% to2.0%, even more preferably between about 1.0% and 2.0% by weight of suchone or more gelling aids over the weight of the shell. In case thegelling aids are cations, the above ranges are expressed as weight ofthe pharmaceutically or food acceptable water soluble salts containingthe cation(s) over the weight of the shell. Exemplary suitable gellingaids' amounts are readily available to a skilled person in the field ofhard capsules manufacturing. For example, it is commonly accepted thatwhen water is about 75% by weight over the weight of the aqueouscomposition, hard capsule shells containing a “target” amount of gellingaids can be obtained via a dip-molding process by using aqueouscompositions containing about ¼ (i.e. 25%) of that target amount(expressed as % by weight over the weight of the composition).

In one embodiment, the aqueous composition of the invention optionallycomprises one or more pharmaceutically or food acceptable dyes and/orcolouring agents.

Said dyes and/or colouring agents may be selected from the groupconsisting of azo-, quinophthalone-, triphenylmethane-, xanthene- orindigoid dyes, iron oxides or hydroxides, titanium dioxide or naturaldyes and mixtures thereof. Further examples are patent blue V, acidbrilliant green BS, red 2G, azorubine, ponceau 4R, amaranth, D+C red 33,D+C red 22, D+C red 26, D+C red 28, D+C yellow 10, yellow 2 G, FD+Cyellow 5, FD+C yellow 6, FD+C red 3, FD+C red 40, FD+C blue 1, FD+C blue2, FD+C green 3, brilliant black BN, carbon black, iron oxide black,iron oxide red, iron oxide yellow, titanium dioxide, riboflavin,carotenes, anthocyanines, turmeric, cochineal extract, clorophyllin,canthaxanthin, caramel, betanin and Candurin® pearlescent pigments.Candurin® is manufactured and marketed by Merck KGaA, Darmstadt, Germanyand consist of titanium dioxide and/or iron oxide—approved food andpharmaceutical colorants in many countries—and potassium aluminiumsilicate as color carrier. The latter is a natural, also widelyapproved, silicate also known under the name of ‘mica’.

In one embodiment, the aqueous composition of the invention contains oneor more pharmaceutically or food acceptable dyes and/or colouring agentsas defined above in an amount suitable to obtain a hard capsule shell asdefined below containing between about 0% and 5.0% by weight, preferablybetween about 0% and 4.0% by weight, more preferably between about 0%and 2.0% by weight of such one or more pharmaceutically or foodacceptable dyes and/or colouring agents over the weight of the shell.Hard capsule shells meeting the above weight % can typically bemanufactured via dip-molding by using aqueous compositions comprisingbetween about 0% and 0.8% by weight, preferably between about 0% and0.6% by weight, more preferably between about 0% and 0.4% by weight ofover the total weight of the aqueous composition. For example, it iscommonly accepted that when water is about 75% by weight over the weightof the aqueous composition, hard capsule shells containing a “target”amount of dyes and/or colouring agents can be obtained via a dip-moldingprocess by using aqueous compositions containing about ¼ (i.e. 25%) ofthat target amount (expressed as % by weight over the weight of thecomposition).

In one embodiment, the aqueous composition of the invention optionallycomprises one or more pharmaceutically or food acceptable sequesteringagents. Preferably, said one or more sequestring agents are selectedfrom the group consisting of EDTA acid, acetic acid, boric acid, citricacid, edetic acid, gluconic acid, lactic acid, phosphoric acid, tartaricacid, or salts thereof, methaphosphates, dihydroxyethylglycine, lecithinor beta cyclodextrin and combinations thereof. Especially preferred areethylenediaminetetraacetic acid, citric acid or any pharmaceutically orfood acceptable salt thereof.

In one embodiment, the aqueous composition of the invention contains oneor more sequestering agents as defined above in an amount suitable toobtain a hard capsule shell as defined below containing between about 0%and 2.0% by weight of such sequestering agents over the weight of theshell. Exemplary suitable sequestering agents' amounts are readilyavailable to a skilled person in the field of hard capsulesmanufacturing. For example, it is commonly accepted that when water isabout 75% by weight over the weight of the aqueous composition, hardcapsule shells containing a “target” amount of sequestering agents canbe obtained via a dip-molding process by using aqueous compositionscontaining about ¼ (i.e. 25%) of that target amount (expressed as % byweight over the weight of the composition).

In one embodiment, the aqueous composition of the invention optionallycomprises one or more pharmaceutically or food acceptable plasticisers.

In one embodiment, the one or more plasticizers are selected in thegroup of plasticizers typically used in the manufacture of hard capsuleshells and in particular in the group consisting of: phtalique esters(e.g. dimethyl-, diethyl-, dibutyl-, diisopropyl- and dioctyl-phtalate);citric esters (e.g. triethyl-, tributyl-, acetyltriethyl- andacetyltributyl-citrate); phosphoric esters (e.g. triethyl-, tricresyl,triphenyl-phosphate); oils (e.g. purified mineral oils, ricin oil, cornoil, cotton oil); butyl stearate; dibutyl sebacate; dibutyl tartrate;diisobutyl adipate, glycerol monostearate; glycerol triacetate;tributyrin; oleic acid; stearic acid; cetylic acid; myristic acid;propylene glycol; glycerol; PEG 4000, PEG 6000, and mixtures thereof.

In one embodiment, and to avoid excessive shell softness, the aqueouscomposition of the invention contains one or more plasticizers asdefined above in an amount suitable to obtain a hard capsule shell asdefined below containing between about 0% and 10% by weight of suchplasticizer(s) over the weight of the shell. Hard capsule shells meetingthe above weight % can typically be manufactured via dip-molding byusing aqueous compositions comprising less than about 2.0%, morepreferably between 0% and 1.0% by weight over the total weight of theaqueous composition.

In one embodiment, the aqueous composition of the invention optionallycomprises one or more pharmaceutically or food acceptable sweetenersand/or flavouring agents.

In one embodiment, the aqueous composition of the invention contains oneor more pharmaceutically or food acceptable sweeteners and/or flavouringagents as defined above in an amount suitable to obtain a hard capsuleshell as defined below containing between about 0% and 1.0% by weight ofsuch sweeteners and/or flavouring agents over the weight of the shell.Exemplary suitable sweeteners and/or flavouring agents' amounts arereadily available to a skilled person in the field of hard capsulesmanufacturing. For example it is commonly accepted that hard capsuleshells containing a “target” amount of sweeteners and/or flavouringagents falling within the ranges identified above can be obtained via adip-molding process by using aqueous compositions containing about ¼(i.e. 25%) of that target amount (expressed as % by weight over theweight of the composition).

In one embodiment, the aqueous composition of the invention optionallycomprises one or more pharmaceutically or food acceptable surfactants.Without wanting to be bound by any theory, it is believed that thesurfactant contributes to the final hard capsule shell surfaceproperties in such a way that the capsule works well on the conventionalautomatic high speed capsule filling equipment. Further guidance onpossible embodiments for the surfactant and the advantages of includinga surfactant in the aqueous composition of the invention can be found inWO2005/105051 filed by the present Applicant.

In one embodiment, the one or more pharmaceutically or food acceptablesurfactants are selected from the group consisting of: sodium laurylsulphate (SLS), dioctyl sodium sulfosuccinate (DSS), benzalkoniumchloride, benzethonium chloride, cetrimide (trimethyltetradecylammoniumbromide), fatty acid sugar esters for example like sorbitol esters (SE)and/or sucrose monolaurate (SML), glyceryl monooleate, polyoxyethylenesorbitan fatty acid esters, polyvinyl alcohol, dimethylpolysiloxan,sorbitan esters or lecithin. In one preferred embodiment, the one ormore pharmaceutically or food acceptable surfactants comprise, morepreferably consist of a mixture of SE and SML.

In one embodiment, the aqueous composition of the invention contains oneor more pharmaceutically or food acceptable surfactants as defined abovein an amount suitable to obtain a hard capsule shell as defined belowcontaining between about 0% and 0.5% by weight of such surfactant(s)over the weight of the shell. Exemplary suitable sweeteners and/orflavouring surfactants' amounts are readily available to a skilledperson in the field of hard capsules manufacturing. For example, it iscommonly accepted that when water is about 75% by weight over the weightof the aqueous composition, hard capsule shells containing a “target”amount of surfactants can be obtained via a dip-molding process by usingaqueous compositions containing about ¼ (i.e. 25%) of that target amount(expressed as % by weight over the weight of the composition).

In one embodiment, the aqueous composition of the invention consists of(i) an aqueous solvent as defined above, (ii) a mono-, di-, andoligosaccharides free pullulan as defined above (iii) a setting systemas defined above and optionally, one or more of the followingingredients: one or more pharmaceutically or food acceptable dyes and/orcolouring agents, one or more pharmaceutically or food acceptablesequestering agents, one or more pharmaceutically or food acceptableplasticisers, pharmaceutically or food acceptable sweeteners and/orflavouring agents and one or more pharmaceutically or food acceptablesurfactants, all as defined above.

In one embodiment, the aqueous composition of the invention consists of(i) an aqueous solvent as defined above, (ii) a mono-, di-, andoligosaccharides free pullulan as defined above (iii) a setting systemas defined above, one or more pharmaceutically or food acceptablesurfactants as defined above and optionally, one or more of thefollowing ingredients: one or more pharmaceutically or food acceptabledyes and/or colouring agents, one or more pharmaceutically or foodacceptable sequestering agents, and one or more pharmaceutically or foodacceptable plasticisers, pharmaceutically or food acceptable sweetenersand/or flavouring agents, all as defined above.

In a further aspect, the present invention relates to a hard capsuleshell comprising (I) moisture, (II) a mono-, di-, and oligosaccharidesfree pullulan and (III) a setting system.

Unless otherwise indicated, in the present invention hard capsules havethe same shape of commercially available, conventional hard capsulesintended for oral administration to a human or animal being. The hardcapsules of the invention can be manufactured by using anon-thermogelling dip molding process and equipment conventionally usedfor the manufacture of hard gelatin capsule shells. As it is disclosedin greater detail below, in the manufacturing process, pin molds(usually kept at about 25° C., i.e. room T) are dipped into an aqueouscomposition containing one or more film-forming polymers, as well as allthe optional ingredients (e.g. setting system(s), plasticizer(s),colouring agent(s) etc) and subsequently withdrawn. The aqueouscomposition is typically kept at T generally comprised between 50° and70° C. The film formed on pins surface is then dried, stripped off thepins and cut to a desired length. Thus, capsules caps and bodies areobtained. Normally, caps and bodies have a side wall, an open end and aclosed end. The length of the side wall of each of said parts isgenerally greater than the capsule diameter. The capsule caps and bodiesare telescopically joined together so as to make their side wallspartially overlap and obtain a hard capsule shell. “Partially overlap”also encompasses an embodiment wherein the side walls of caps and bodieshave substantially the same length so that, when a cap and a body aretelescopically joined, the side wall of said cap encases the entire sidewall of said body. Unless otherwise indicated, “capsule” refers tofilled capsule shells whereas “shell” specifically refers to an emptycapsule. Since the hard capsule shells of the invention can be filledwith substances in liquid form, it is intended that if desired the hardcapsules of the invention may be sealed or banded according toconventional techniques. Also, the hard capsule shells of the inventioncan be manufactures so to have a specific capsule shell design thatprovides with certain advantages as the possibility to pre-lock emptycaps and bodies and complete the filling step in a differentlocation/time. An example of advantageous designs suitable to beimplemented in the hard capsule shells of the present invention can befound in WO2009/138920 (specifically when capsule filling is a liquidsubstance) or WO2009/050646.

Exemplary and preferred amounts of pullulan, setting system, dyes and/orcolouring agents, sequestring agents, plasticizers, sweeteners and/orflavouring agents and surfactants in the shell of the invention areindicated above in the context of the aqueous composition.

Unless otherwise indicated, when discussing about shells and capsules ofthe invention, moisture and water are terms that can be usedinterchangeably.

The hard capsule shells of the invention typical comprise between 2% and20%, preferably between about 5% and 15% by weight of moisture over thetotal weight of the shell.

Typically, a capsule of the invention is dried to an LOD comprisedbetween about 10% and 15% can be considered acceptable for a capsuleshell of the invention. However, LOD can be brought to lower % by e.g.further drying, capsule shell storage at low RH or following capsuleshell filling with hygroscopic substances.

In one embodiment, the hard capsule shell of the invention consists of(I) moisture as defined above, (II) a mono-, di-, and oligosaccharidesfree pullulan as defined above (III) a setting system as defined aboveand optionally, one or more of the following ingredients: one or morepharmaceutically or food acceptable dyes and/or colouring agents, one ormore pharmaceutically or food acceptable sequestering agents, one ormore pharmaceutically or food acceptable plasticisers, pharmaceuticallyor food acceptable sweeteners and/or flavouring agents and one or morepharmaceutically or food acceptable surfactants, all as defined above.The presence of one or more pharmaceutically or food acceptablesurfactants is preferred.

In one embodiment, the shells of the inventions may be externally coatedwith a suitable coating agent like cellulose acetate phthalate,polyvinyl acetate phthalate, methacrylic acid gelatins, hypromellosephthalate, hydroxypropylmethyl cellulose phthalate, hydroxyalkyl methylcellulose phthalates, hydroxypropyl methylcellulose acetate succinate ormixtures thereof to provide e.g. enteric properties.

In one embodiment, the hard capsule shell of the invention is a shellobtainable using the aqueous composition of the invention as disclosedabove. In one embodiment, the hard capsule shell of the invention is ashell obtainable using the aqueous composition of the invention asdisclosed above and a dip-molding process of the invention as disclosedbelow.

In one embodiment, the hard capsule shell of the invention has a shellthickness (after drying till a shell LOD of lower than about 10%) lowerthan about 250 μm, preferably lower than about 150 μm, more preferablygreater than about 70 μm, even more preferably between about 70 and 150μm, even more preferably of about 100 μm. These shell thickness valuescannot be obtained with non dip-molding manufacturing methods (e.g.injection molding) that typically give thicker shells of about 300 to500 μm.

With respect to currently commercially available pullulan capsule shells(e.g. NPcaps® from Capsugel, France) the shells of the invention presentimproved characteristics in terms of shell mechanical propertiesespecially at low shell LOD levels, and an equally good or improvedshell dissolution profile, shell transparency, capsule surface staticand gliding properties, absence of unpleasant smell, capsulecompatibility with automatic high speed capsule filling equipment.

In one embodiment, low shell LOD levels preferably mean a shell LODlower than about 9%, preferably lower than about 8%, more preferablycomprised between about 7 and 8%. This LOD value is commonly adoptedwhen filling shells with moisture sensitive or hygroscopic materials sothat a lower shell brittleness at these value represent a significantadvantage.

In one embodiment, shell mechanical properties at a specific shell LODare tested according to the so-called “tube test”. This test is wellknown to skilled men working in the field of hard capsules and oneprocedure for its performance is also disclosed in the literature (D.Cadé and N. Madit, “Liquid Filling in Hard Gelatin Capsules—PreliminarySteps”, Bulletin Technique Gattefossé, 1996). During the tests, the % ofbroken capsule shells in a sample of several tens of shells is evaluatedby varying the shell LOD: the lower the %, the lower the shellbrittleness, the better the shell mechanical properties. As shown in theexamples of this application, the capsule shells of the presentinvention showed a statistically reduced brittleness with respect toconventional pullulan capsules at shell LOD levels lower than about 10%,and notably at shell LOD levels comprised between about 7 and 8%.

In one embodiment, shell dissolution profile is tested by filling asample of 6 hard capsule shells with acetaminophen, and verifying if oneor more of the filled capsules in said batch release less than about 80%of the filled acetaminophen at 45 minutes in simulated gastric fluid(pepsin) after 6 months at storage conditions of 40° C. and 75% RH.Acetaminophen dissolution is evaluated in accordance with, and usingequipment and method conditions disclosed in USP-32 for Acetaminophen.Such “Dissolution test”, using an immediate release reference (i.e.acetaminophen), indicates changes in the capsule dissolution rate due toan inappropriate dissolution properties of the pullulan used for shellmanufacture. In one embodiment, the method to test dissolutionproperties is performed as disclosed in D. Cadé and N. Madit, “LiquidFilling in Hard Gelatin Capsules—Preliminary Steps”, Bulletin TechniqueGattefossé, 1996.

In one embodiment, shell transmittance is tested according to thefollowing procedure: cut a 1 cm² flat film sample from a capsule(size#2, natural transparent body), then measure its transmittance at25° C. by using SHIMADSU UV1600PC spectrophotometer from 400 nm to 700nm with 2 nm pitches of continuous scanning.

In one embodiment, shell static and glide properties are testedaccording to the methods disclosed in WO03/039522 (“Surface coatedcapsules”), Examples 2 and 1, respectively.

In one embodiment, shell smell is tested according to the followingprocedure: approximately 200 capsules are put in a 200 mL flask thenplugged. Comparative capsules are put in another flask and plugged aswell. Flasks are put in an oven at 40° C. for about 1 hour. Thenpanellists compare smell strength and rank it according to a scalecomprising five smell strength levels ranging from 1 (lowest smell) to 5(strongest smell).

In one embodiment, capsule compatibility with automatic high speedcapsule filling equipment is tested according to the followingprocedure: in capsule filling machine (CFM), a vacuum pressure of about20 cm Hg is applied to separate pre-locked bodies and caps in a batch of1500 capsules. If all capsules can be separated, the test is re-run witha pressure of about 10 cm Hg. The number of capsules that cannot beseparated at this step is recorded. Other parameters that are monitoredare the number of capsules that incur cracking or deforming duringseparation. The highest the number of capsules that separate withoutdefects, the higher the compatibility with the CFM machine.

In a further aspect, the present invention relates to a hard capsulecomprising a shell as defined above filled with one or more substancesin solid, semi-solid and/or liquid form.

In one embodiment, the one or more substances to be filled into theshells of the invention are substances that are sensitive to moisture orsubstances that are hygroscopic.

Unless otherwise indicated, substances that are sensitive to moisturepreferably means substances that undergo any known chemical degradationfollowing the contact with even minor amounts of water such as moisturecontent typical of hard gelatin capsules (for example, shell LODcomprised between 12% and 25%).

In one embodiment, preferred substances to be filled into the shells ofthe invention are selected from the group consisting of: placentapowder, aloe extract powder, pollen, activated carbon, glucosamine,carnitine, alfa-lipoic acid, royal jelly, propolis extract, garlic,seaweed extract, mineral salts, sesame extract, coenzyme, chitin,chitosan, chondroitin, glutathione, asteroid extract, soybean powder,lecithin, tea or herb powders, corn silk powder, lactobacillus, berry,wheat albumin, arginine, angleworm powder, ginger powder, champignonpowders, kiwi extract, carrot extract, hyaluronic acid, grape-seedextract, collagen extract, lycopene, turmeric extract, lutein, catechin,plant extracts, nucleic acid, sugars, chlorella, vitamin, astaxanthin,yeast, shark extract, shellfish extract, other antioxidation compounds,and mixtures thereof.

In one embodiment, the hard capsules of the invention can be madetamper-proof by using appropriate techniques to make the joint betweencapsule caps and bodies permanent. Typically, sealing or bandingtechniques can be used where these techniques are well-known to anyskilled person in the field of hard capsules. In this connection, it isconventional practice to perform banding and/or sealing using polymersolutions in water/ethanol or water/isopropanol solutions. Traces ofsuch non water solvents thus can be found if an elemental analysis isperformed on a sealed or banded capsule of the invention without makinga distinction between ingredients that are part of the shell andingredients that are part of the band or sealing subsequently applied.

In a further aspect, the present invention relates to a dip-moldingmethod for the manufacture of hard capsule shells as defined above, saidprocess comprising the steps of:

(a) dipping pins into an aqueous composition as defined above

(b) withdrawing the dipping pins from the aqueous composition and

(c) drying the composition adhered on the dipping pins so as to obtain ashell;

wherein the steps (a) to (c) are performed in the order in which theyare presented.

Dip-molding processes for the manufacture of hard capsules using cooledpins and a solution of one or more film-forming polymers and optionallyone or more gelling agents (e.g. carrageenans) and/or co-gelling agents(e.g. inorganic cations) is known since decades. For patent reviewdisclosing this process one can see e.g. U.S. Pat. No. 5,264,223, U.S.Pat. No. 5,756,123 and U.S. Pat. No. 5,756,123 (all relating to HPMCcapsule shells and entailing the use of setting systems).

In one embodiment, the method of the present invention comprises beforestep (a), a step of providing an aqueous composition as defined above.

In one embodiment, the method of the present invention comprises beforestep (a) but after the step of providing an aqueous composition asdefined above, a further step of defoaming the aqueous compositionprovided.

In one embodiment, the method of the present invention comprises betweensteps (b) and (c), a further step of turning pins from a “top-down”dipping position (position of step (a)) to a “top-up” drying position(position in step (c)). In this additional step the pins are rotatedabout a horizontal axis of about 180° with respect to the dippingposition of step (a).

In one embodiment, the pins in step (a) are kept at a pin dippingtemperature which is preferably comprised between about 20° C. and 30°C., more preferably comprised between about 20° C. and 25° C.

In one embodiment, the aqueous composition in step (a) are kept at anaqueous composition dipping temperature which is preferably comprisedbetween about 45° C. and 65° C., more preferably between about 50° C.and 65° C.

In one embodiment, step (c) of drying is performed according toconventional drying techniques typically applied in the field of hardcapsules and by using conventional equipment known to the skilled personfor this purpose. In one embodiment, step (c) of drying can be performedaccording to any technique commonly known for this purpose, for exampleby placing the pins in conventional ovens.

The method of the presence invention allows maintaining capsuledimension during production.

In one embodiment, maintenance of capsule dimension is tested accordingto the following in-line procedure: during manufacturing, an operatortakes a sample of a predefined number of capsules and measures capsulebodies and caps weight as well as side and top wall thickness.

In a further aspect, the present invention relates to the use of amono-, di-, and oligosaccharides free pullulan for the manufacture ofhard capsule shells.

Further embodiments and advantages of the present invention will becomeapparent to a skilled reader in light of the examples provided below.Unless otherwise specified, all parts and percentages are by weight.Composition viscosities were determined by Brookfield viscometer.

Example 1

Hard capsule shell manufacturing—Samples 1-3 were prepared according tothe following general procedure. Pullulan powder (amount to have a 22.5%by weight over the weight of the final aqueous composition—see below forpullulan grade details) is mixed with a gelling agent (see below forsetting system definition and relative amount in each sample). To 5 kgof deionized water under stirring at room temperature a gelling aid isadded (see below for gelling aid definition and relative amount in eachsample), followed by addition of the above mixture. The powder additionand stirring speeds were very high in order to avoid the forming oflumps. The solution is heated up to 70° C. under stirring to totallydissolve the gelling agent and pullulan. It is possible to dissolve thecomponents directly at 70° C., but the tendency of pullulan to lump ismuch higher. The pullulan solution thus prepared is defoamed under slowstirring and then poured into a dipping dish of a pilot machine ofconventional hard gelatin capsule production equipment. Processconditions: as for standard NPcaps® capsules manufacturing, i.e. dishtemperature 60° C.; mold pin temperature 37° C.—Standard dippingprofile.

-   Sample 1: Size#2, natural, transparent, hard capsule shells—mono-,    di-, and oligosaccharides amount 1.1% w/w—pullulan solution    viscosity of 1470 mPa·s at 60° C.—gelling agent (carrageenan) 0.3%    by weight over shell weight—co-gelling agent (KCl) 1.65% w/w by    weight over shell weight;-   Sample 2: Size#2, natural, transparent, hard capsule shells—mono-,    di-, and oligosaccharides amount 2.4% w/w—pullulan solution    viscosity of 1250 mPa·s at 60° C.—setting system as for Sample 1;-   Sample 3 (comparative): Size#2, natural, transparent, standard    NPcaps® capsules (from Capsugel, France) manufactured with    commercial grade Pullulan PI-20—setting system as for Sample 1.

Brittleness of Samples 1-3 was tested by Tube Test after storage atdifferent RH conditions:

Broken capsules Sample # 14% RH 23% RH 33% RH 1 100 4 0 2 100 0 0 3(comparative) 100 70 14

Compared to standard NPcaps® capsules manufactured with commercial gradeof Pullulan PI-20, a significant reduction of brittleness (level ofbroken capsules) was observed for storage under 33 & 23% RH.

Example 2

-   Sample 4 (comparative): Size#2, natural, transparent, pullulan    capsules manufactured with commercial grade Pullulan PI-10—setting    system as for Sample 1—manufacturing process as for Sample 1 but    pullulan concentration in the aqueous composition was raised from    20% to 30% w/w.

Brittleness of Samples 3 and 4 (both comparative) was tested by TubeTest after storage at different shell moisture contents (obtained bystoring shells at different RH conditions):

% Broken capsules 12% moisture 10% moisture 8% moisture Sample # contentcontent content 3 (comparative) 0  5 40 4 (comparative) 0 20 60

Compared to standard NPcaps, capsules manufactured with pullulan PI-10showed a significant increase in brittleness (level of broken capsules)when stored to achieve lower moisture contents. This example incombination with Example 1 shows that the level of mono-, di, andoligosaccharides in the pullulan is critical to obtain capsules with animproved brittleness whereas the average molecular weight of pullulanchains is not.

Example 3

-   Sample 5: natural, transparent, hard capsule shells—mono-, di-, and    oligosaccharides amount 0.5% w/w—pullulan batch dimension is 400 Kg    (industrial scale)—pullulan source is corn starch—manufacturing    process and setting system as for Sample 1;-   Sample 6: repeat of sample 6

Brittleness of Samples 3 and 5-6 was tested by Tube Test after storageat different RH conditions:

% Broken capsules Sample # 14% RH 23% RH 33% RH 3 (comparative) 100 7014 5 100 18 0 6 100 10 0

Example 3 confirms reproducibility and repeatability of positive resultswith an industrial-scale batch of pullulan from corn starch on a pilotmachine of conventional hard gelatin capsule production equipment.

Example 4

Hard capsule shells of Samples 7 and 8 were manufactured under standardproduction conditions i.e. with industrial full size hard capsulesmanufacturing machines following the general procedure disclosed in“Multiparticulate Oral Drug Delivery, edited by Isaac Ghebre-SellassieDrugs and the Pharmaceutical Sciences Vol. 65 Marcel Dekker, Inc.1994—Chapter 14, Capsule Shell composition and Manufacturing RonnieMillender p. 357-383”. The aqueous composition was optimized to themanufacture of pullulan capsules and consisted of:

Pullulan 22.5% by weight over weight of the composition,

Gelling agent (carrageenan) 0.067% by weight over weight of thecomposition

Co-gelling agent (KCl) 0.371% by weight over weight of the composition

Water to 100%

-   Sample 7: Capsule Lot#166181—Size #2, natural transparent hard    capsule shells—mono-, di-, and oligosaccharides amount 0.5%    w/w—pullulan source was non-GMO corn starch;-   Sample 8 (comparative): Capsule Lot#165651—conventional NPcaps®    manufactured using Pullulan PI-20 commercial grade

Brittleness of Samples 7 and 8 was tested by Tube Test after storage atdifferent RH conditions:

% Broken capsules Sample # 11% RH 22% RH 50% RH 7  2 0 0 8 (comparative)63 5 0

Despite the formulation optimization, the gap in terms of brittlenessafter storage at low RH between conventional pullulan grade capsules andthe capsules of the present invention is remarkable.

Example 5

-   Sample 9: Capsule Lot#175451—Size #2, natural transparent hard    capsule shells—mono-, di-, and oligosaccharides amount 0.5%    w/w—pullulan source was tapioca starch—manufacturing process and    setting system were as for Example 4;

Brittleness of Samples 7 and 9 was tested by Tube Test after storage atdifferent RH conditions:

% Broken capsules Sample # 11% RH 22% RH 50% RH 7 2 0 0 9 5 0 0

Example 5 shows that pullulan capsules of the invention from Tapiocastarch are equivalent in terms of mechanical properties under lowmoisture conditions to pullulan capsules of the invention from non-GMOcorn starch.

Example 6

-   Sample 10: Lot#180531—Size #2, natural transparent hard capsule    shells—mono-, di-, and oligosaccharides amount 0.9% w/w—pullulan    batch #0A2921—pullulan source was tapioca starch—process conditions    and setting system were as for Example 4;-   Sample 11: Lot#180541—Size #2, white opaque body and cap pullulan    hard capsule shells—mono-, di-, and oligosaccharides amount 0.9%    w/w—pullulan batch #0A2921—pullulan source was tapioca    starch—process conditions were as follows: TiO₂ was dispersed in an    aqueous solution, then poured into a pullulan aqueous solution to    obtain 2.0% w/w TiO₂ pigment content in the finished capsule.    Capsule shells were manufactured according to the method and using    the setting system described in Example 4;-   Sample 12: Lot#180981—Size #2, natural transparent hard capsule    shells—mono-, di-, and oligosaccharides amount 2.4% w/w—pullulan    batch #0B0321—pullulan source was tapioca starch—process conditions    and setting system were as for Example 4;-   Sample 13: Lot#180521—Size #2, white opaque body and cap pullulan    hard capsule shells—mono-, di-, and oligosaccharides amount 2.4%    w/w—pullulan batch #0B0321—pullulan source was tapioca    starch—process conditions and setting system were as for sample 11;

Brittleness of Samples 8-13 was tested by Tube Test after 1 week storageat 11% RH:

Sample # % Broken capsules  9  8 10 10 11 33 12 15 13 55 8 (comparative)80

This example shows (i) a positive effect on capsule shell brittleness byreducing the content of mono-, di-, and oligosaccharides (ii) inclusionof TiO₂ as opacifier entails a slight worsening in shell brittleness.Thus, in case opaque capsules are desired, it is preferred to use lowamounts of mono-, di-, and oligosaccharides amounts such as lower thanabout 1.0% w/w.

Example 7

-   Sample 14: Lot#179801—Size #2, natural transparent hard capsule    shells—Conventional NPcaps® manufactured using commercial grade    Pullulan PI-20—process conditions and setting system were as for    Example 4;    3 batches (approximately 50 capsules, each) of natural, transparent    pullulan capsules (one batch of Sample 9, one batch of Sample 10 and    one batch of Sample 14) were filled with the following hygroscopic    products:

Garlic Extract from Musyu-NinNiku powders PS-II Osada Co. LTD; initialpowder moisture content: 1.3% w/w

Bilberry extract from Bilbelon-25 from Tokiwa Phytochemical Co., LTD;initial powder moisture content: 1.8% w/w

Garcinia extract from Garcitrin (Garcinol 0.5%) Sabinsa Japan Co., LTD;initial powder moisture content: 2.1% w/w

After 1 week storage at room conditions (20° C., 50% RH) capsulesmoisture was controlled:

Capsule LOD % after 1 week storage Sample # Capsule initial LOD % GarlicBilberry Garcinia  9 11.6 6.5 7.1 6.7 10 11.4 6.3 6.8 6.8 14(comparative) 12.0 6.5 7.0 7.3

As predictable, moisture exchange between the hygroscopic fill and thecapsule shell resulted in capsules dehydration. After moisture control,the filled capsules were tested according to the tube-test:

% Broken capsules Sample # Garlic Bilberry Garcinia  9 0 0 0 10 0 0 0 14(comparative) 12  0 4

This example further confirms the advantage of the capsules of thepresent invention in terms of brittleness at an LOD of about 7%.

Example 8

-   Sample 15: Lot#93181—Size #2, white opaque body and cap pullulan    hard capsule shells—Conventional NPcaps® manufactured using    commercial grade Pullulan PI-20—process conditions and setting    system were as for sample 11;    2 batches (approximately 50 capsules, each) of white opaque pullulan    capsules (one batch of Sample 11 and one batch of Sample 15) were    filled with Garlic and Garcinia extracts (see Example 7 for    details):

After 1 week storage at room conditions (20° C., 50% RH) capsules weretested with tube test:

% Broken capsules Sample # Garlic Garcinia 11  4 0 15 (comparative) 80 8

A significant improvement in terms of brittleness between standardcommercial pullulan capsules and the capsules of the present inventionis apparent.

Example 9

Capsule smell was measured by a group of 5 panellists on capsules fromSample 7 and Sample 8:

Score for each panellist # Sample # #1 #2 #3 #4 #5 Avg. Dev. Tot. 7 3 33 3 3 3 0 15 8 3 3 2 3 3 2.8 0.4 14 (comparative)

No significant difference of smell was detected between pullulancapsules of the invention and NPcaps pullulan commercial capsules.

Example 10

Following the test procedure disclosed in the present application,compatibility with CFM (capsule filling machines) was evaluated forcapsule from samples 7 and 8:

Capsules cracking − Capsule batch = Non separation at 10 cm Hg − Sample# 1000 capsules Capsule batch = 1500 capsules 7 0 0 8 0 0 (comparative)

This example shows that shell compatibility with modern filling machinesis substantially identical for the hard capsule shells of the presentinvention and conventional pullulan hard capsule shells.

The invention claimed is:
 1. A hard capsule shell comprising a settingsystem, and at least one pullulan, wherein the pullulan is substantiallyfree of mono-, di-, and oligosaccharides, wherein the amount of mono-,di-, and oligosaccharides in the pullulan is less than 2.5% by weight ofpullulan on a dry solid basis, and wherein the shell comprises between65% and 99% of the pullulan by weight of the shell.
 2. The hard capsuleshell according to claim 1, wherein the pullulan has an averagemolecular weight ranging from about 100 KDa to about 400 KDa.
 3. Thehard capsule shell according to claim 1, wherein the pullulan has a meltviscosity at about 60° C. ranging from about 500 cPs to about 1500 cPs.4. The hard capsule shell according to claim 1, wherein the pullulancomprises less than about 1.0% of mono-, di- and oligosaccharides byweight of pullulan, on a dry solid basis.
 5. The hard capsule shellaccording to claim 1, wherein the pullulan is produced from corn ortapioca starch.
 6. The hard capsule shell according to claim 1, furthercomprising at least one ingredient selected from the group consisting ofpharmaceutically or food acceptable dyes and/or colouring agents,pharmaceutically or food acceptable sequestering agents,pharmaceutically or food acceptable plasticisers, pharmaceutically orfood acceptable sweeteners and/or flavouring agents, pharmaceutically orfood acceptable surfactants, and mixtures thereof.
 7. A hard capsulecomprising the hard capsule shell according to claim 1 filled with oneor more substances, wherein at least one substance is in solid,semi-solid and/or liquid form.
 8. The hard capsule shell according toclaim 1, wherein the shell has a shell thickness less than about 500 μm.9. The hard capsule shell according to claim 8, wherein the shell has ashell thickness less than about 250 μm.
 10. The hard capsule shellaccording to claim 1, wherein the pullulan comprises less than about0.7% of mono-, di-, and oligosaccharides by weight of pullulan, on a drysolid basis.
 11. The hard capsule shell according to claim 1, whereinthe pullulan comprises less than about 0.6% of mono-, di-, andoligosaccharides by weight of pullulan, on a dry solid basis.